Plasma Microparticle Tissue Factor Activity in Patients With Antiphospholipid Antibodies With and Without Clinical Complications

Thrombosis Research (Impact Factor: 2.45). 12/2013; 133(2). DOI: 10.1016/j.thromres.2013.11.027
Source: PubMed


Antiphospholipid syndrome (APS) is defined by the association of autoantibodies to certain phospholipid-binding proteins with arterial or venous thrombosis ('AT' or 'VT', respectively), and/or pregnancy-related morbidity (PM). Antiphospholipid antibodies (aPLA) promote activation of several cell types including monocytes, resulting in procoagulant tissue factor (TF) expression that may contribute to the vascular complications. Since TF synthesis by monocytes is frequently accompanied by release of TF-bearing microparticles, we hypothesized that plasma microparticle TF activity (MP-TF) may be elevated in APS patients and contribute to thrombosis and/or PM. Platelet-poor plasma specimens were obtained from 30 patients with definite APS and 72 patients with asymptomatic aPLA from the Antiphospholipid Syndrome Collaborative Registry (APSCORE). MP-TF was measured by an in-house factor Xa generation assay. The two groups were well matched for gender, age, ethnicity, proportions with underlying SLE, and aPLA profiles. MP-TF (median and (IQR)) in asymptomatic aPLA subjects was 0.09pg/mL (0.05-0.14) compared to 0.13pg/mL (0.10-0.17) in APS (p<0.001). No differences in MP-TF levels were observed between APS subjects with PM, thrombosis, or PM+thrombosis. Similarly, among subjects with either APS or asymptomatic aPLA, MP-TF did not differ in the presence or absence of underlying SLE. Prospective studies will be required to determine if plasma MP-TF activity is causally related to thrombotic or gestational complications in APS.

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    • "We used the to-date most widely applied and best validated assay for plasma MP-TF activity determination in the present study. MP-TF activity has been measured with this assay in cancer patients with acute VTE [7], in a large cohort of cancer patients that were prospectively followed for VTE [22], in pancreatic cancer patients with different stages-and histologic grades of disease [23], in patients with antiphospholipid antibodies [24] and in plasma after stimulation of monocytes with lipopolysaccharide [25]. Lee et al. assessed the lower detection limit of the MP-TF activity assay used in the present study [26]. "
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    ABSTRACT: Background: Tissue factor (TF) is the main in-vivo initiator of blood coagulation. Microparticles (MPs) are small procoagulant membrane vesicles. Elevated TF-bearing MPs have been found in different prothrombotic conditions and MP-associated TF activity may contribute to the pathogenesis of unprovoked deep vein thrombosis (DVT). Objective: To determine MP-TF activity levels at diagnosis of DVT and at four additional time points during the course of one year in a well-defined group of patients with unprovoked DVT of the lower limb. Patients/Methods: In this study, 41 patients with acute unilateral symptomatic and unprovoked DVT of the lower limb were included and followed for 1 year. Venous blood samples for determination of MP-TF activity were drawn at diagnosis of acute DVT, and 1-, 3-, 6-, and 12 months later. In addition, 10 young and healthy control subjects were included. Results: The median MP-TF activity was 0.06 pg/mL (25th-75th percentile: 0.0-0.53) in patients with acute DVT and 0.18 pg/mL (0.07-0.33) in healthy controls, and did not differ significantly (p = 0.35). No significant changes in MP-TF activity were found in the follow-up measurements. MP-TF activity did also not differ significantly between patients with proximal-or distal DVT and between those with-or without residual DVT after 6 months. Conclusions: MP-TF activity is low at the acute event in patients with unprovoked DVT of the lower limb and remains unchanged during the course of the disease. Our data do not support the hypothesis that TF-bearing MPs play a determining role in the pathogenesis of unprovoked DVT.
    Thrombosis Research 08/2014; 134(5). DOI:10.1016/j.thromres.2014.07.041 · 2.45 Impact Factor
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    ABSTRACT: The antiphospholipid syndrome is characterized by venous or arterial thrombosis and/or recurrent fetal loss in the presence of circulating antiphospholipid antibodies. These antibodies cause activation of endothelial and other cell types leading to the release of microparticles with procoagulant and pro-inflammatory properties. The aims of this study were to characterize the levels of endothelial cell, monocyte, platelet derived, and tissue factor-bearing microparticles in patients with antiphospholipid antibodies, to determine the association of circulating microparticles with anticardiolipin and anti-β2-glycoprotein antibodies, and to define the cellular origin of microparticles that express tissue factor. Microparticle content within citrated blood from 47 patients with antiphospholipid antibodies and 144 healthy controls was analyzed within 2 hours of venipuncture. Levels of Annexin-V, CD105 and CD144 (endothelial derived), CD41 (platelet derived) and tissue factor positive microparticles were significantly higher in patients than controls. Though levels of CD14 (monocyte-derived) microparticles in patient plasma were not significantly increased, increased levels of CD14 and tissue factor positive microparticles were observed in patients. Levels of microparticles that stained for CD105 and CD144 showed a positive correlation with IgG (R = 0.60, p = 0.006) and IgM anti-beta2-glycoprotein I antibodies (R = 0.58, p = 0.006). The elevation of endothelial and platelet derived microparticles in patients with APS and their correlation with anti-β2-glycoprotein I antibodies suggests a chronic state of vascular cell activation in these individuals and an important role for β2-glycoprotein I in development of the pro-thrombotic state associated with antiphospholipid antibodies.
    Thrombosis Research 11/2014; 135(1). DOI:10.1016/j.thromres.2014.11.011 · 2.45 Impact Factor
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    ABSTRACT: Elevated levels of endothelial cell-derived extracellular vesicles (EV) circulate in patients with antiphospholipid antibodies (APLA), and APLA, particularly those against β2-glycoprotein I (β2GPI), stimulate EV release from endothelial cells. However, the effects of endothelial cell-derived EV have not been characterized. To determine the mechanism by which EV released from endothelial cells by anti-β2GPI antibodies activate unstimulated endothelial cells. We used IL-1 receptor inhibitors, siRNA to Toll-like receptors, and microRNA (miRNA) profiling to assess mechanism(s) by which EV released from endothelial cells exposed to anti-β2GPI antibodies activated unstimulated endothelial cells. Anti-β2GPI antibodies caused formation of an endothelial cell inflammasome and release of EV that were enriched in mature IL-1β, had a distinct miRNA profile, and caused endothelial activation. However, activation was not inhibited by an IL-1β antibody, IL-1 receptor antagonist or IL-1 receptor siRNA. Endothelial cell activation by EV required IRAK4 phosphorylation and was inhibited by pretreatment of cells with TLR7 siRNA or RNase A, which degrades single-stranded RNA. Profiling of miRNA in EV released from endothelial cells incubated with β2GPI and either control IgG or anti-β2GPI antibodies revealed numerous differences in the content of specific miRNAs, including a significant decrease in mIR126. These observations demonstrate that although anti-β2GPI-derived endothelial EV contain IL-1β, they activate unstimulated endothelial cells through a TLR7 and single-stranded RNA dependent pathway. Alterations in miRNA content may contribute to the ability of EV derived from endothelial cells exposed to anti-β2GPI antibodies to activate unstimulated endothelial cells in a paracrine manner. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.
    Journal of Thrombosis and Haemostasis 08/2015; 13(10). DOI:10.1111/jth.13072 · 5.72 Impact Factor
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